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研究生: 楊廷毅
Yang, Ting-Yi
論文名稱: 邏輯演繹序列串聯質譜法應用於牡蠣肝醣多醣之結構鑑定
Logically Derived Sequence Tandem Mass Spectrometry for Structural Determination of Polysaccharide-Oyster Glycogen
指導教授: 倪其焜
Ni, Chi-Kung
陳頌方
Chen, Sung-Fang
口試委員: 倪其焜
Ni, Chi-Kung
陳頌方
Chen, Sung-Fang
張煥正
Chang, Huan-Cheng
口試日期: 2024/07/17
學位類別: 碩士
Master
系所名稱: 化學系
Department of Chemistry
論文出版年: 2024
畢業學年度: 112
語文別: 中文
論文頁數: 107
中文關鍵詞: 牡蠣肝醣多糖結構鑑定邏輯演繹序列串聯質譜法
英文關鍵詞: oyster glycogen, polysaccharide, structure identification, LODES/MSn
研究方法: 實驗設計法
DOI URL: http://doi.org/10.6345/NTNU202401463
論文種類: 學術論文
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  • 傳統的多醣分析方法包含許多複雜的步驟,例如衍生化、泛甲基化、氣相層析質譜法以及核磁共振光譜(nuclear magnetic resonance, NMR)。我們實驗室近期開發出一套新的質譜方法,邏輯演繹序列串聯質譜法(logically derived sequence tandem mass spectrometry, LODES/MSn),用於醣的結構鑑定。本研究中,多醣被水解成單醣、雙醣、和各種不同尺寸的寡醣,再使用高效率液相層析法及邏輯演繹序列串聯質譜法鑑定這些單醣、雙醣和寡醣。本次研究我們利用邏輯演繹序列串聯質譜法分析牡蠣肝醣的水解產物之結構,決定了牡蠣肝醣的最小分支結構,幫助了解牡蠣肝醣結構裡的實際分支狀況。邏輯演繹序列串聯質譜法能以極少量的樣品決定醣類的結構,並縮短鑑定所需的時間,將是鑑定多醣基本結構的有力工具。

    Conventional polysaccharide analysis involves complex procedures, including chemical derivatization, permethylation, gas chromatography/mass spectrometry, and nuclear magnetic resonance spectrometry. We have recently developed a new mass spectrometry method, Logically Derived Sequence Tandem Mass Spectrometry (LODES/MSn), for the structural identification of carbohydrates. In this study, polysaccharides were hydrolyzed into monosaccharides, disaccharides, and various sizes of oligosaccharides. These monosaccharides, disaccharides, and oligosaccharides were identified by using high-performance liquid chromatography and LODES/MSn. In this research, we utilized LODES/MSn to analyze the structure of hydrolysis product of oyster glycogen, determining the minimal branching structures within oyster glycogen, which aids in understanding real branching patterns in oyster glycogen. LODES/MSn allows for the determination of carbohydrate structures with small amounts of sample and significantly reduces the time required for identification, making it a powerful tool for identifying the basic structures of polysaccharides.

    1 第一章 緒論 1 1.1 醣類的介紹 1 1.2 醣類的結構鑑定方式 4 1.3 研究背景及動機 4 2 第二章 實驗方法 6 2.1 實驗藥品、器材與儀器設備 6 2.1.1 實驗藥品 6 2.1.2 實驗儀器 6 2.2 實驗流程 7 2.3 實驗步驟 8 2.3.1 實驗藥品前處理 8 2.3.2 多醣水解 8 2.3.3 固相萃取(solid phase extraction, SPE) 10 2.3.4 粒徑篩析層析法(size exclusion chromatography, SEC) 11 2.3.5 高效液相層析(high performance liquid chromatography, HPLC) 11 2.3.6 質譜儀(Mass spectrometry) 12 3 第三章 邏輯演繹序列串聯質譜法 13 4 第四章 實驗結果與討論 21 4.1 單醣分析 21 4.2 雙醣分析 25 4.3 三醣分析 36 4.4 四醣分析 53 4.5 五醣分析 89 5 第五章 結論 104 6 參考文獻 105

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